Compositions and methods for combination pharmacological treatments to induce a prolonged, mild decrease in core body temperature

ABSTRACT

The present invention is directed to combination pharmacological treatments that induce a prolonged, modest decrease in core body temperature, to the range of −95° F. to −98.5° F. (−35° C. to −36.7° C.) to mitigate and delay the early onset and progression of fatal disease processes associated with premature aging, such as those characteristic of progeria (Hutchinson-Gilford Progeria Syndrome, HGPS), Wiedemann-Rautenstrauch syndrome (neonatal progeroid syndrome), Werner syndrome, and other rare disorders characterized by premature aging, and to prolong the shortened lifespan of children afflicted with these diseases. Short-term chronic administration of the pharmaceutical treatments to a person suffering from mild concussion may also have value in ameliorating neurological damage/symptoms associated with mild concussion. Chronic administration of the pharmacological treatments may also have value in normal human populations or in high-risk human populations. Similar benefits may be realized in other mammals such as canines, felines and other non-human species.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/224,725 filed Aug. 1, 2016, which claims benefit under 35 U.S.C. §119(e) of U.S. Patent Application No. 62/200,004, filed Aug. 1, 2015.Both applications are hereby incorporated herein by reference in theirentireties.

BACKGROUND TO THE INVENTION

Hypothermia

The Baltimore Longitudinal Study of Aging (BLSA)

In the Baltimore Longitudinal Study of Aging (BLSA), men with core bodytemperatures below the median lived significantly longer than men withbody temperatures above the median, in the absence of caloricrestriction (CR). The BLSA has accumulated almost 30 years of follow-updata in a large population and continues today (Roth et al. (2002)Science 297, 811).

Specifically, in the largest reported longitudinal study of bodytemperature in humans, it was reported, a decade ago, in men in the BLSAstudy that participants with core body temperatures in the lower 50% ofthe study population had significantly lower mortality than those withbody temperatures in the upper 50% over 25 years of follow-up (Soare etal. (2011) Aging 3, 374-379).

In humans the average internal temperature is generally considered to be˜37.0° C. (−98.6° F.), although it varies among individuals. Roth et al.examined the effects of various longevity-related markers from the BLSA.Consistent with the beneficial effects of caloric restriction on agingand lifespan in other animals, men with lower temperatures had greatersurvival than those with higher temperatures, but the BLSA men were notcalorie-restricted.

Furthermore, lower body temperature was reported to be one of only threeindependent factors influencing longevity (FIG. 1).

The Scripps Research Institute Study

In a 2006 study, scientists at the Scripps Research Institute found thatreducing the core body temperature of mice extended their medianlifespan, by up to 20% (Conti et al. (2006) Science 314, 825-828). Thiswas the first time that changes in body temperature had been shown toaffect lifespan in warm-blooded animals. The study demonstrated that itwas possible to increase lifespan in mice by modest, but prolonged,lowering of the core body temperature and that the longer lifespan wasachieved independent of caloric restriction.

Prior to this study, researchers had known that core body temperatureand aging were related in cold-blooded animals. Scientists had alsoknown that lifespan could be extended in warm-blooded animals byreducing the number of calories they consumed, which also lowered corebody temperature as a consequence. However, the degree of caloricrestriction needed to extend lifespan is not easy to achieve, even inmice.

Prior to the Scripps study, key questions about the relationship betweencaloric restriction, core body temperature, and lifespan remainedunanswered. Was caloric restriction itself responsible for longerlifespan, with reduced body temperature simply a consequence? Or was thereduction of core body temperature a key contributor to the beneficialeffects of caloric restriction?

Mice in the Scripps study were allowed to eat as much food as theywished, and the experimental and control mice ate the same amountsuggesting that hypothermia—and not caloric restriction—may be moredirectly tied to longevity (Conti et al. (2006) Science 314, 825-828).Other examples illustrating and supporting this point follow.

The Naked Mole Rat

Another example of a lower body temperature's effects on aging is thenaked mole rat (Heterocephalus glaber), a subterranean, extraordinarilylong-lived eusocial mammal (Kim et al. (2011) Nature 479, 223-227).Although the size of a mouse, the naked mole rat's maximum lifespanexceeds 30 years and makes this animal by far the longest living rodent(˜10 times the typical lifespan of comparably sized rodents).

Naked mole rats show negligible senescence, no age-related increase inmortality, and high fecundity until death. In addition to delayed aging,naked mole rats are resistant to both spontaneous cancer andexperimentally induced tumorigenesis. They also maintain healthyvascular function longer in their lifespan than shorter-lived rats. Thenaked mole rat's normal core body temperature is a full 6° C. lower thanany other rodent.

It was at first considered that the naked mole rat's exceptionallifespan was associated with its burrowing lifestyle that includedintermittent periods of ‘feast’ and prolonged ‘famine’ (calorierestriction). However, the precise reason for their longevity is stilldebated and is likely the result of many factors.

Longevity was thought to be related to the naked mole rat's ability tosubstantially reduce its metabolism during hard times, and so to preventaging-induced damage from oxidative stress. However, when naked molerats were removed to captivity, they survived to their full potentialand remained resistant to disease whether fed their native diet in afeast-famine pattern or a full ad libitum diet, ingested at will. Theircore body temperatures, however, remained low. As noted with the Scrippsstudy, the suggestion was that hypothermia—and not caloricrestriction—may be more directly tied to the naked mole rat's resistanceto disease and remarkable longevity.

Progeria and Progeria-Like Diseases in Children

Progeria, also known as Hutchinson-Gilford Progeria Syndrome (HGPS), isa rare, fatal genetic condition of accelerated aging in children (Gordonet al. (2012) J. Cell Biol. 199, 9-13). All children with progeria dieof the same heart disease that affects millions of normally aging adults(arteriosclerosis), but instead of this occurring at 60-70 years of age,the children may suffer strokes and heart attacks even before age 10.

Progeria affects ˜1 in 4-8 million newborns. There are an estimated200-250 children living with Progeria worldwide at any one time. Itaffects both sexes equally and all races (Sarkar & Shinton (2001)Postgrad. Med. J. 77, 312-317).

Progeria is a progressive genetic disorder that causes children to agerapidly, beginning in their first 2 years of life. Children withprogeria generally appear normal at birth. By 12 months, signs andsymptoms, such as slow growth and hair loss, begin to appear. Theaverage life expectancy for a child with progeria is ˜13, but some withthe disease die younger and some live 20 years or longer.

Heart problems or strokes are the eventual cause of death in mostchildren with progeria. There is currently no treatment for thecondition.

Researchers have discovered a single gene mutation responsible for HGPS.The gene is known as lamin A (LMNA), which encodes a protein necessaryfor holding the nucleus of a cell together. It is believed that thegenetic mutation renders cells unstable, which appears to lead toprogeria's characteristic aging process (Gordon et al. (2012) J. CellBiol. 199, 9-13; Navid et al. (2012) Bioinformation 8, 221-224).

Children with progeria usually develop severe hardening of the arteries.This is a condition in which the walls of their arteries stiffen andthicken, often restricting blood flow. Most children with progeria dieof complications related to atherosclerosis, including problems with theblood vessels that supply the heart (cardiovascular problems), resultingin heart attack and congestive heart failure, and problems with theblood vessels that supply the brain (cerebrovascular problems),resulting in stroke (Gerhard-Herman et al. (2012) Hypertension 59, 92-7;Low et al. (2005) BMC Medical Genetics 6, 38).

There are other progeroid syndromes that run in families (Navarro et al.(2006) Hum. Mol. Genet. 15, R151-161). They include, but are not limitedto, Wiedemann-Rautenstrauch syndrome (WRS), Werner syndrome (WS), BloomSyndrome, and Cockayne syndrome (CS). In Wiedemann-Rautenstrauchsyndrome, also known as neonatal progeroid syndrome, the onset of agingbegins in the womb, and signs and symptoms are already apparent atbirth. Werner syndrome typically begins in adolescence or earlyadulthood.

These inherited progeroid syndromes also cause rapid aging and shortenedlife span. In the case of Werner syndrome, it has a global incidencerate of <1 in 100,000 live births, although the incidence in Japan andSardinia is higher, where it affects ˜1 in 20,000-40,000 and ˜1 in50,000, respectively. In 2006, there were ˜1,300 reported cases of WSworldwide.

Hypothermia and its Relationship with Aging

In recent years, many factors have been theorized to prevent or delaythe onset of diseases that affect longevity or to have a direct,independent, or multi-factor inter-related effect on increasinglifespan. Influences such as free radicals, caloric restriction, loweredbody temperature, and even components in red wine (e.g., resveratrol)have each been explored.

Even when links have been suggested or established, there remains thepractical issue of how to effectively achieve decreased caloric intake,free radical reduction, or the benefits of red wine, for example,without significant lifestyle modifications that must be sustainedlong-term at often extreme levels and with great difficulty andinconsistency in implementation.

In the case of body temperature reduction, there is presently nopractical means of achieving mild and controlled hypothermia (such as adecrease from normal baseline to the range of ˜95° F. to ˜98° F.) thatmay be of clinical benefit on a chronic-use basis. In fact, studies haveshown that a lower average core body temperature by ˜0.2° C., whichsounds like a modest reduction, is statistically significant and similarto the reduction observed in long-lived, calorie-restricted mice (Soareet al. (2011) Aging, 3, 374-379).

Although moderate hypothermia induction (a ˜2-4° C. reduction in coretemperature) has been established as having clinical benefit, forexample in the prevention of organ and neurological damage post-cardiacarrest resuscitation, current methods of achieving hypothermic inductionare relatively extreme acute interventions that currently can beaccomplished only through the application of artificial ‘forced’ orexternal methods (e.g., cold blankets, iced saline infusion) accompaniedby extreme procedures and processes that, for example, requireintubation and ventilatory assistance, heavy sedation and inducedparalysis simply to offset the body's natural defense of shivering inresponse to external attempts to reduce core body temperature.

The ‘mechanical’ or ‘forced’ methodologies in current use are clearlyimpractical for chronic, routine application and long-term induction ofa controlled, regulated reduction in body temperature.

Much evidence of lowered core body temperature's independent effect onprolonging lifespan has emerged in recent years. These findingsdemonstrate the importance of a reduction of body temperature inmodulating longevity.

In the case of the Scripps study, scientists lowered core bodytemperature directly, without restricting food intake. In cold-bloodedanimals, such as roundworms (C. elegans) and fruit flies (Drosophila),this task is straightforward; core body temperature can be loweredsimply by changing the temperature of the environment.

However, for warm-blooded animals, the task is much more challenging.The Scripps experiment focused on the preoptic area of the hypothalamus,a structure in the brain that acts as the body's thermostat and iscrucial to temperature regulation. Just as holding something warm nearthe thermostat in a room can fool it into thinking that the entire roomis hotter so that the air conditioning turns on, the Scripps Researchteam reasoned that they could reset the brain's thermostat by producingheat nearby. To do so, they created a mouse model that produced largequantities of uncoupling protein 2 in hypocretin neurons in the lateralhypothalamus, near the preoptic area. The action of uncoupling protein 2produced heat, which diffuses to other brain structures, including thepreoptic area. Indeed, the extra heat worked to induce a continuousreduction of the core body temperature of the mice, lowering it from˜0.3 to ˜0.5° C. The scientists were then able to measure the effect oflowered core body temperature on lifespan, finding that the mice withlowered core body temperature had significantly longer median lifespanthan those that did not have a lowered core body temperature.

The researchers performed several experiments to make sure that otherfactors were not contributing to the lowered core body temperature. Theyconfirmed that the experimental mice were normal in their ability togenerate a fever, and that these mice moved around to about the samedegree as normal mice. Additionally, the researchers verified that thehypocretin neurons producing uncoupling protein 2 were not involved intemperature regulation. Importantly, the mice in this study were allowedto eat as much food as they wished, and the experimental and controlmice ate the same amount (Conti et al. (2006) Science 314, 825-828).

Several other studies have documented a lowering of core bodytemperature by caloric restriction in mice, rats, and rhesus monkeys.Interestingly, however, ad libitum fed transgenic mice overexpressingthe uncoupling protein 2 in hypocretin neurons (Hcrt-UCP2) also had alower core body temperature and a 16% greater life expectancy thanwild-type animals, independent of caloric intake.

Although body temperature is recognized as a clinically usefulphysiological parameter in the context of infection or extremeenvironmental exposures, few epidemiological studies have included bodytemperature as a routine measurement. One consistent observation thathas emerged from studies of human body temperature, however, has beenthat advanced age is associated with lower body temperature.

In the landmark cross-sectional studies of Wunderlich in the 1860s—whichincluded 25,000 participants and established the “normal” bodytemperature of 98.6° F.—lower body temperatures were observed in elderlyparticipants (Mackowiak et al. (1992) JAMA 268, 1578-1580).

Therapeutic Hypothermia

Hypothermia, as opposed to a modest and controlled reduction in normalbody temperature, is a condition in temperature-regulating organismswhere the core body temperature is reduced below the normal range.Hypothermia has been used clinically for more than 40 years to protectbodily organs from various pathophysiological insults, includingischemic insults such as cardiac arrest, hemorrhage, hypergravity, andhypoglycemia, and to reduce the toxicity of various drugs andenvironmental toxicants (Gordon (2001) Ernerg. Med. J. 18, 81-9).

While the precise mechanisms responsible for the therapeutic effects ofhypothermia are not fully understood, hypothermia causes a generalreduction in cellular metabolism (Polderman (2008) Lancet 371, 1955-69).This reduction during hypothermia is especially beneficial to highlyaerobic organs, such as the brain and heart, under ischemic conditionsbecause it leads to reduced demand for oxygen.

Forced hypothermic methods are used for therapeutic hypothermia. Forcedhypothermia involves the use of external mechanical and/or endovascularcooling methods to extract heat from the body to reduce the bodytemperature below the normal temperature set point. External coolingmethods include immersing a subject in a cool bath or applying blanketsor pads with cooled water circulating through channels in the walls ofthe blanket or pad to the skin of a subject. Other external methodsinclude wetting of the skin or hair of the subject, cooling the airaround the subject, and blowing air across the subject's skin.Endovascular cooling generally involves the rapid intravenousadministration of an iced saline solution or heat exchange with aspecially designed catheter.

Hypothermia after Cardiac Arrest

In heart surgery, induction of hypothermia (to 28-32° C.) before cardiacarrest has been used successfully since the 1950s to protect the brainagainst global ischemia.

Successful use of therapeutic hypothermia after cardiac arrest in humanswas also described in the late 1950s (Benson et al. (1959) Anesth.Analg. 38, 423-428; Williams & Spencer (1958) Ann. Surg. 148, 462-468;Ravitch et al. (1961) N. Engl. J. Med. 264, 36-38) but was subsequentlyabandoned because of uncertain benefit and difficulties with its use(Marion et al. (1996) Crit. Care Med. 24, 81S-89S).

More recently, the induction of hypothermia after the return ofspontaneous circulation (ROSC) has been associated with improvedfunctional recovery and reduced cerebral histological deficits invarious animal models of cardiac arrest (Horn et al. (1991) ActaNeuropathol. (Berl). 81, 443-449; Sterz et al. (1991) Crit. Care Med.19, 379-389; D'Cruz et al. (2002) J. Cereb. Blood Flow Metab. 22,848-851; Hicks et al. (2000) J. Cereb. Blood Flow Metab. 20, 520-530).Further preliminary human studies were conducted subsequently (Bernardet al. (1997) Ann. Emerg. Med. 30, 146-153, Sanada et al. (1998) Masui.47, 742-745, Yanagawa et al. (1998) Resuscitation 39, 61-66, Nagao etal. (2000) J. Am. Coll. Cardiol. 36, 776-783, Zeiner et al. (2000)Stroke 31, 86-94, Felberg et al. (2001) Circulation 104, 1799-1804,Hachimi-Idrissi et al. (2001) Resuscitation 51, 275-281, Callaway et al.(2002) Resuscitation 52, 159-165).

At the time of publication of the American HeartAssociation/International Liaison Committee on Resuscitation (AHA/ILCOR)2000 Guidelines for Cardiopulmonary Resuscitation and EmergencyCardiovascular Care, the evidence was insufficient to recommend use oftherapeutic hypothermia after resuscitation from cardiac arrest(AHA/ILCOR (2000) Resuscitation 46, 1-447), but two subsequent studies,one conducted in post-cardiac arrest resuscitated patients in Australia(Bernard et at. (2002) NEJM 346, 557-563) and the second in post-cardiacarrest resuscitated patients conducted by the Hypothermia After CardiacArrest Study Group (HACA) in nine European centers (HACA (2002) NEJM346, 549-556, became the basis for the AHA/ILCOR recommendation for theuse of therapeutic hypothermia in the treatment of post cardiac arrestresuscitated patients issued in 2003.

-   -   Indeed, the AHA/ILCOR Recommendations, issued in June/July 2003,        state that “On the basis of the published evidence to date, the        ILCOR ALS Task Force has made the following recommendations:        Unconscious adult patients with spontaneous circulation after        out-of-hospital cardiac arrest should be cooled to 32° C. to        34° C. for 12 to 24 hours when the initial rhythm was VF        [ventricular fibrillation]. Such cooling may also be beneficial        for other rhythms or in-hospital cardiac arrest.”

Thus, the use of therapeutic hypothermia is now accepted as a standardintervention for the preservation of vital organ systems in theaftermath of cardiac arrest (AHA/ILCOR recommendations).

Therapeutic hypothermia can be achieved mechanically, using medicaldevices such as, for example, the ThermoSuit®. The ThermoSuit® cools thepatient with direct water on skin contact. The water flows over thepatient and is returned to the pump. A continuous flow of water iscycled through drawing heat off the body in a much more rapid processthan either gel pads or cooling blankets; a treatment period of ˜30 minis usually sufficient to cause a body core temperature reduction of ˜3°C. The suit is portable and is used with a conventional gurney. As thepatient reaches the target temperature the water is drawn off thepatient and the ThermoSuit® is removed.

While the application of therapeutic hypothermia is accepted as thestandard intervention for the preservation of vital organ function inthe aftermath of cardiac arrest (AHA/ILCOR recommendation), there is atpresent no effective pharmacological means to lower and maintain bodytemperature.

Chronotherapeutics

The existence of circadian rhythms in cardiovascular disease iswell-established. It is also known that heart rate and blood pressurenormally peak during the morning hours and reach a nadir in the lateevening, around bedtime. The incidence of myocardial infarction, stroke,sudden cardiac death, and myocardial ischemia increases during theearly-morning hours. Angina attacks occur in a diurnal cycle; theiroccurrence is common in the hours shortly after an individual beginsactivity, after waking. Body temperature also follows circadianpatterns, where core temperature is typically lower in the morning hoursand higher in the afternoon and evening hours.

Based on these relationships, researchers have begun to apply thescience of chronotherapeutics, or the timing of drug effect withbiologic need, to improve cardiovascular outcomes. Traditional treatmentregimens for conditions associated with circadian variation typically donot account for circadian fluctuations in disease activity.

Chronotherapeutic regimens are intended to provide pharmacologicalintervention at the most appropriate time point(s), in accordance withcircadian rhythms. The concept of chronotherapeutics in treatingcardiovascular diseases includes dosing traditional agents at specifictimes throughout the day, the development of new agents, and thedevelopment of chronotherapeutic formulations and combinations of drugswith special release mechanisms targeted at inducing the greatest effectduring the pre- and post-waking morning surge in heart rate (HR) andblood pressure (BP) and/or, in select populations, during sleep.

Chronotherapeutic agents are specifically intended to provide peakplasma concentrations when their effect is most needed (e.g., in theearly morning hours with regard to HR and BP; in the case of regulatingbody temperature, the reverse may be true). Further, the lowestconcentrations of drugs typically occur at night, when HR and BP aretypically lowest and, consequently, cardiovascular events are leastlikely to occur. However, special consideration also needs to be givento overnight levels of the drug(s) for nocturnal hypertensives, and forall hypertensives in the pre- and post-waking hours, where HR and BP(and the calculated RPP, a measure of myocardial oxygen demand and asurrogate predictor of target organ damage and cardiovascular events)rise.

Pharmacological Means for Inducing Therapeutic Hypothermia

There is presently no effective pharmacological means to lower andmaintain body temperature over a prolonged period of time. Therapeutichypothermia is currently restricted to short-term use in acute medicalsituations that require the induction of mild hypothermia (bodytemperature in the range of ˜32° C. to ˜34° C.).

For example, US 2012/0282227 A1 (Katz) describes compositions andmethods for inducing moderate hypothermia in a subject as an acute,short-term intravenously delivered intervention targeting a therapeuticrange of between 32 and 34° C. for 12-24 h duration. However, this isnot suitable for chronic, long-term application.

Such acute, mild hypothermia is distinct from modest and controlledhypothermia (a decrease from normal baseline to the range of ˜95° F. to˜98° F.), which may be of clinical benefit on a chronic-use basis anddoes not require intravenous administration. The compositions describedby Katz are reported to be useful in treating acute clinical insults,including, but not limited to, cerebral ischemic insults, such aspost-cardiac arrest resuscitation neurological damage, stroke, spinalcord injury, or traumatic brain injury.

The Katz patent application (US 2012/0282227) also mentions the use of aneurotensin analog, NT69L, for inducing moderate hypothermia.Neurotensin is a 13-amino-acid neuropeptide that regulates the releaseof leuteinizing hormone and prolactin. An analog of neurotensin refersto a polypeptide analog of neurotensin that may have an amino acidsequence that is longer, shorter, or the same length as the amino acidsequence of neurotensin. Neurotensin analogs may includenon-naturally-occurring amino acids and may also include non-amino acidcompounds. However, NT69L has hypothermic tolerance issues that wouldprevent its chronic use.

Methods are also reported to be useful for maintaining regulated mildhypothermia in a subject for a prolonged period of time (i.e., 12-24 hor more) and for reducing the time necessary to induce regulatedhypothermia, as compared to mechanical methods of induction (US2012/0282227).

The compositions or multidrug combinations of Katz′ invention comprise aregulated hypothermic compound or a dopamine receptor agonist, avasoactive compound, and an anti-arrhythmic compound or a serotonin5-HT₃ receptor antagonist.

As an example, the patent application describes

-   -   A composition comprising:    -   a regulated hypothermic compound or a dopamine receptor agonist,    -   a vasoactive compound, and    -   an anti-arrhythmic compound or a serotonin 5-HT₃ receptor        antagonist.

Various combinations of drug components are described. In some examples,the compositions comprise ethanol and optionally at least one of avasoactive compound, an anti-hyperglycemic compound, a dopamine receptoragonist, an anti-arrhythmic compound, a serotonin 5-HT₃ receptorantagonist, an anti-oxidant, a vitamin, and N-acetylcysteine.

In the examples where the composition or multidrug combination comprisesethanol and optionally additional agents, the method of administrationrequires a two-phase method of delivery of the composition. Theinvention described further recognizes that a two phase delivery, adelivery of a high concentration of the multidrug combination followedby delivery of a lower concentration of the multidrug combination. Thistwo phase method of delivery may be accomplished by a rapid infusion ofthe composition to induce hypothermia followed by a period of slowinfusion, rapidly reducing the body temperature of a patient andmaintaining the hypothermic state for a sustained period of time. Usingthis method of delivery, the composition may comprise a regulatedhypothermic compound or a dopamine receptor agonist, a vasoactivecompound, and an anti-arrhythmic compound or a serotonin 5-HT₃ receptorantagonist and additional additives, as discussed above.

Additionally, using the two-phase method of delivery, the compositionmay comprise ethanol and at least one of a vasoactive compound, ananti-arrhythmic compound, a serotonin 5-HT₃ receptor antagonist, ananti-oxidant, a vitamin, N-acetylcysteine, and an anti-hyperglycemiccompound. This two-phase delivery method can be used to deliver any ofthe compositions of the invention and provides significant benefits to apatient.

The methods disclosed include a method for inducing hypothermia in asubject comprising administering to the subject a multidrug combinationcomprising a regulated hypothermic compound or a dopamine receptoragonist, a vasoactive compound, and an anti-arrhythmic compound or aserotonin 5-HT₃ receptor antagonist.

In those embodiments where the clinical insult is of an acute nature,the methods were capable of preventing or limiting permanent injuries orstress, particularly if used within the first few hours after theclinical insult. The methods were reported to be useful in treatingpatients suffering from brain injuries, heart injuries, kidney injuries,cardiac bypass patients, patients suffering cardiac arrest, patientswith neurologic injury, infants with hypoxic-ischemic encephalopathy,and injuries of vital organs related to ischemia-reperfusion. Thus,these methods can be used to reduce the stress and discomfort ofshivering and to positively benefit and prevent injuries to the brain,heart, kidneys, and other organs. The methods lead to a reduction inmortality or a reduction in adverse effects attributed directly orindirectly to the clinical event.

Studies have shown that a lower average core body temperature by ˜0.2°C., which is a modest reduction, is statistically significant andsimilar to the reduction observed in long-lived, calorie-restricted mice(see Soare et al. (2011) Aging 3, 374-379). In the case of bodytemperature reduction, there is presently no practical means ofachieving and sustaining such modest and controlled hypothermia (adecrease from normal baseline to the range of ˜95° F. to ˜98° F.) thatmay be of clinical benefit on a chronic-use basis.

Thus, there is a continuing need to provide drugs or drug products thatsafely lower body temperature by BLSA study amounts (˜95° F. to ˜98°F.), suitable for chronic low-dose use.

Embodiments of the present invention address that continuing need,providing effective pharmacological means to achieve modest andcontrolled hypothermia (a decrease from normal baseline to the range of˜95° F. to ˜98° F.).

SUMMARY OF THE INVENTION

The present invention is directed to combination pharmacologicaltreatments that induce a prolonged modest decrease in core bodytemperature, into the range of ˜95° F. to ˜98.5° F. (˜35° C. to ˜36.7°C.) to mitigate and delay the early onset and progression of fataldisease processes associated with premature aging, such as thosecharacteristic of progeria (also known as Hutchinson-Gilford ProgeriaSyndrome or HGPS), Wiedemann-Rautenstrauch syndrome (also known asneonatal progeroid syndrome), Werner syndrome, and other rare disorderscharacterized by premature aging, and to prolong the shortened lifespanof children afflicted with these diseases.

Short-term chronic administration of the pharmaceutical treatment to aperson suffering from mild concussion (e.g., sports-related injury,automobile accident, falls, bumps) may also have value in amelioratingthe neurological damage/symptoms associated with mild concussion where amodest decrease in body temperature for a prescribed number of days orshort course of treatment could minimize damage and potentially hastenrecovery time.

Chronic administration of the pharmacological treatment may also havevalue in normal human populations or in high-risk human populations,such as those with a family history of heart disease or cancer or inpatients with compromised cardiac status or organ damage, by delaying orpreventing the onset and progression of potentially fatal,life-shortening disease processes and prolonging the potentiallyshortened lifespan of at-risk patients or the potential life span ofhuman populations, whether or not at risk. Similar benefits may berealized in other mammal populations, such as canines, felines, andother non-human species.

The present invention is directed to drugs or drug products, including,but not limited to, combinations of drugs, that safely lower bodytemperature by BLSA study-like amounts (lower 50^(th) percentile),suitable for chronic low-dose use, providing an effective method toachieve modest and controlled hypothermia (a decrease from normalbaseline into the range of ˜95° F. to ˜98° F.).

The focus of the present invention is on the pharmacological inductionof prolonged modestly reduced body temperature (in the range of ˜95° F.to ˜98° F.) to achieve the presumed organ protective and resultinglifespan-extending effects of hypothermia.

Embodiments of the invention include unique combination drugformulations (two or more medications in one drug product) at doseslower than those normally used in monotherapy with the drugs inquestion.

Embodiments of the invention include unique combination drugformulations (two or more pharmaceutical agents in one drug product) atdoses similar to those normally used in monotherapy with the drugs inquestion.

Embodiments of the invention include unique combination drugformulations (two or more pharmaceutical agents in one drug product) atdoses higher than those normally used in monotherapy with the drugs inquestion.

Embodiments of the invention include combination drug productsformulated as time-release or extended-release products.

Similar to the discussion above on cardiovascular disease, circadianfluctuations in disease activity, and chronotherapeutics, there is someevidence, for example, that olanzapine may be optimally effective in the‘dark period’ with respect to temperature decrease. Metformin may beideally suited for its desired effects occurring in the ‘light period.’Thus, embodiments of the present invention include chronotherapeuticformulations of drugs or drug products, including combinations of drugs,that safely lower body temperature by BLSA study-like amounts (lower50^(th) percentile), suitable for chronic low-dose use, providingeffective pharmacological methods to achieve modest and controlledhypothermia (a decrease from normal baseline into the range of ˜95° F.to ˜98° F.).

Embodiments of the invention include combination drug productsformulated as chronotherapeutic products.

Embodiments of the invention include a pharmaceutical composition andmethod of treatment where the combination pharmaceutical entitycontaining prescribed doses of an atypical anti-psychotic drug withhypothermia-inducing properties, such as olanzapine (Evers andScheurink, Dutch Research Consortium, 2011) or risperidone (Cope et al.(2009) Physiol. Behay. 96, 457-463), or an indole alkaloidanti-psychotic and anti-hypertensive drug, such as reserpine, and anoral anti-diabetic drug in the biguanide class, such as metformin (Wu etal. (2008) JAMA 299, 185-93)), which has caloric-restriction and otherdesired effects, such as weight loss, reduced insulin resistance, andincreased metabolism (e.g., Ji et al. (2013) PloS One 8, e57222), tobalance or offset the potential for weight gain, decreased metabolism,and increased insulin resistance associated with atypicalanti-psychotics, or similarly, an anti-inflammatory, anti-allergicimmunomodulator, such as amlexanox, that has been shown to reverseobesity, diabetes, and fatty liver in mice (Reilly et al. 2013 Nat. Med.19, 313-321), administered orally, for example, in the form of a pill,gel-cap, liquid, cream, or spray form.

While the primary indication for use of atypical anti-psychotics is thetreatment of schizophrenia, drugs in this class have a documentedhistory of off-label use to treat anxiety disorder, attention deficithyperactivity disorder, dementia and severe geriatric agitation, majordepressive disorder, eating disorders, insomnia, obsessive compulsivedisorder (OCD), post-traumatic stress disorder (PTSD), personalitydisorders, substance abuse, and Tourette's syndrome (Maglione et al.,Off-label Use of Atypical Anti-psychotics, Comparative EffectivenessReviews, No. 43, Agency for Healthcare Research and Quality (US), DHHS,Report No.: 11-EHC087-EF, September 2011), and that in the normal courseof development of all FDA approved drugs, including atypicalanti-psychotics such as olanzapine, safety trials in healthy normalhuman subject volunteers are also required, and this has been the casefor all previously approved subject drugs referenced here. While notapproved for a specific indication other than schizophrenia, the use ofthese drugs in normal volunteers under strict regulatory scrutiny iswell-established. Relative to the current application, it should benoted that the use of approved drugs to elicit a potentially positive‘side effect’ and apply it for that purpose to the treatment of asecondary medical indication distinct from the approved use(repurposing) is not unprecedented. In a classic example, minoxidil, anapproved anti-hypertensive, was repurposed for the treatment of malepattern baldness and alopecia on the basis of an observation of anincidental minoxidil side-effect, hair growth, which resulted indevelopment of the topical form of minoxidil, known as Rogaine®.

While a modest, sustained decrease in body temperature is the focus ofthe current invention, and the primary purpose of the inclusion ofmetformin is to offset the potential negative effects of thehypothermia-inducing drug component, metformin may have complementaryindependent properties that may expand or enhance the value of thedesired effect of the combination. For example, Mayo Clinic researchersfound that women with ovarian cancer who took metformin were, onaverage, nearly four times more likely to survive for 5 years afterdiagnosis than women not taking the drug (Kumar et al. (2013) Cancer119, 555-562). Other studies suggest that metformin helps slow or stoptumor growth in other cancers too (Hassan et al. (2010) Cancer 116,1938-1946; Yang (2011) Front. Med. 5, 115-117; Kirpichnikov et al.(2002) Ann Intern Med. 137, 25-33). To further support that metforminmay have complementary independent properties that may supplement orenhance the primary body temperature lowering focus of the invention,there is growing evidence that metformin reduces the risk ofcardiovascular disease. For example, it was reported by Dr. Peter Boyle(International Prevention Research Institute, Lyon, France) that in 29different studies, metformin was associated with a 10% reduction in therisk of cardiovascular events, with the drug associated with a 10%reduction in the risk of heart failure in seven studies and a 12%reduction in the risk of MI in nine studies (Boyle, P. et al. Safety ofglucose-lowering medications: The Diabetes Adverse Event Monitor(DIABAMON) project: II cardiovascular disease. American DiabetesAssociation 2013 Scientific sessions; Jun. 23, 2013; Chicago, Ill.Abstract 1405-P).

In a preferred embodiment, the pharmaceutical composition isadministered to children afflicted with progeria or other diseasescharacterized by premature aging and shortened life spans, as a means toslow the onset and progression of associated age-related diseases andeffectively extend the expected lifespan.

In a preferred embodiment, the pharmaceutical composition isadministered to a person suffering from mild-to-moderate concussion, toameliorate the neurological damage/symptoms associated with concussion(e.g., sports-related injury, minor automobile accident, falls, bumps)where a modest decrease in body temperature for a prescribed number ofdays or short course of treatment could minimize damage and potentiallyhasten recovery time.

In a preferred embodiment, the pharmaceutical composition isadministered to a person with the intent to prolong longevity.

In a preferred embodiment, the pharmaceutical composition isadministered to a menopausal female to alleviate, reduce the occurrenceof or minimize the effect of hot flashes.

In a preferred embodiment, the olanzapine or risperidone or reserpinecomponent of the combination exerts a body temperature-lowering effect.

Embodiments of the invention include pharmaceutical combinations ofolanzapine or risperidone or reserpine with metformin

Embodiments of the invention include pharmaceutical combinations ofolanzapine or risperidone or reserpine with amlexanox.

Embodiments of the combination drug invention may include apharmaceutical component with temperature reduction properties, such asa neurotensin analog, such as T69L (Katz patent application US2012/0282227) or similar analogs, in combination with, for example,metformin or amlexanox.

Embodiments of the combination drug invention may include apharmaceutical component with body temperature-reducing properties suchas β-blockers, clonidine, ethanol, phenothiazines, neuroleptics andsedative hypnotics, with metformin or amlexanox.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Correlations between longevity and body temperature, insulinlevels, and DHEAS levels.

Correlations between longevity and three factors—body temperature,insulin levels, and DHEAS levels—were found both in monkey studies andin the Baltimore Longitudinal Study of Aging (BLSA). The figuresummarizes these findings. For male rhesus monkeys, it graphs theeffects of caloric restriction versus control monkeys who were notcalorie-restricted (parts A, B, and C). For men in the BLSA, it showsthe effects of the same three factors on natural longevity, expressed interms of survival time (parts D, E, and F). Men who had lower bodytemperatures and lower insulin levels—although they were not caloricallyrestricted, as the rhesus monkeys were—lived longer (top survival curvesin the graphs) than those who had higher values in those parameters(bottom survival curves). Source: Roth et al. (2002) Science 297, 811.

FIG. 2. Survival and mortality curves.

Mice were fed ad libitum on sterilized breeder chow. Median life spanwas (A) 20% (females) and (C) 12% (males) greater in Hcrt-UCP2 micerelative to wild-type littermates. Complementary logarithmic plots (Band D) suggest that the ratio of the hazard rates for Hcrt-UCP2 andwild-type littermates was approximately constant with time. Thisassessment was verified by testing the significance of age as atime-dependent covariate. tg, transgenic; wt, wild-type. Source: Contiet al. (2006) Science 314, 825-828.

FIG. 3. Mean body temperature by age and gender.

Source: Waalen & Buxbaum (2011) J. Gerontol. A Biol. Sci. Med. Sci. 66A,487-492.

DETAILED DESCRIPTION OF THE INVENTION

As stated above, and described in detail below, the present invention isdirected to drugs or drug products, providing effective pharmacologicalmeans to achieve modest and controlled hypothermia (a decrease fromnormal baseline to the range of ˜95° F. to ˜98° F.).

The present invention is directed to combination pharmacologicaltreatments that induce a prolonged modest decrease in core bodytemperature, into the range of ˜95° F. to ˜98.5° F. (˜35° C. to ˜36.7°C.), to mitigate and delay the early onset and progression of fataldisease processes associated with premature aging, such as thosecharacteristic of progeria (also known as Hutchinson-Gilford ProgeriaSyndrome or HGPS), Wiedemann-Rautenstrauch syndrome (also known asneonatal progeroid syndrome), Werner syndrome, and other rare disorderscharacterized by premature aging, and to prolong the shortened lifespanof children afflicted with these diseases.

Short-term chronic administration of the pharmaceutical treatment to aperson suffering from mild concussion (e.g., sports-related injury,automobile accident, falls, bumps) may also have value in amelioratingthe neurological damage/symptoms associated with mild concussion, wherea modest decrease in body temperature for a prescribed number of days orshort course of treatment could minimize damage and potentially hastenrecovery time.

Chronic administration of the pharmacological treatment may also havevalue in normal human populations or in high-risk human populations,such as those with a family history of heart disease or cancer or inpatients with compromised cardiac status or organ damage, by delaying orpreventing the onset and progression of potentially fatal,life-shortening disease processes and prolonging the potentiallyshortened lifespan of at-risk patients or the potential life span ofhuman populations, whether or not at risk. Similar benefits may berealized in other mammalian populations such as canines, felines, andother non-human species.

The present invention is directed to drugs or drug products, includingcombinations of drugs, that safely lower body temperature by BLSAstudy-like amounts (lower 50^(th) percentile), suitable for chroniclow-dose use, providing effective pharmacological means to achieve mildand controlled hypothermia (a decrease from normal baseline to the rangeof ˜95° F. to ˜98° F.).

The focus of the present invention is on the pharmacological inductionof prolonged, modestly reduced body temperature (in the range of ˜95° F.to ˜98° F.) to achieve the presumed organ-protective and resultinglifespan-extending effects of hypothermia.

Embodiments of the invention include unique combination drugformulations (two or more medications in one drug product) at doseslower than those normally used in monotherapy with the drugs inquestion.

Embodiments of the invention include unique combination drugformulations (two or more medications in one drug product) at dosessimilar to those normally used in monotherapy with the drugs inquestion.

Embodiments of the invention include unique combination drugformulations (two or more medications in one drug product) at doseshigher than those normally used in monotherapy with the drugs inquestion.

Embodiments of the invention include combination drug productsformulated as time-release or extended-release products.

Embodiments of the invention include combination drug productsformulated as chronotherapeutic products.

Embodiments of the invention include a pharmaceutical composition andmethod of treatment where the combination pharmaceutical entitycontaining prescribed doses of an atypical anti-psychotic drug withhypothermia-inducing properties, such as olanzapine (Evers andScheurink, Dutch Research Consortium, 2011) or risperidone (Cope et al.(2009) Physiol. Behay. 96, 457-463), or an indole alkaloidanti-psychotic and anti-hypertensive drug, such as reserpine, and anoral anti-diabetic drug in the biguanide class, such as metformin (Wu etal. (2008) JAMA 299, 185-93)), which has caloric-restriction and otherdesired effects, such as weight loss, reduced insulin resistance, andincreased metabolism (e.g., Ji et al. (2013) PloS One 8, e57222), tobalance or offset the potential for weight gain, decreased metabolism,and increased insulin resistance associated with atypicalanti-psychotics, or similarly, an anti-inflammatory, anti-allergicimmunomodulatory drug, such as amlexanox, that has been shown to reverseobesity, diabetes, and fatty liver in mice (Reilly et al. (2013) Nat.Med. 19, 313-321), administered orally, for example, in the form of apill or in liquid or spray form, or as a drug-impregnated patch.

In a preferred embodiment, the pharmaceutical composition isadministered to children afflicted with progeria or other diseasescharacterized by premature aging and shortened life span.

In a preferred embodiment, the pharmaceutical composition isadministered to a person suffering from concussion, to ameliorate theneurological damage/symptoms associated with concussion (e.g.,sports-related injuries, automobile accidents, falls, bumps).

In a preferred embodiment, the pharmaceutical composition isadministered to a person suffering from or at risk of developing cardiacdisease, such as atherosclerotic disease, stroke, or having a familyhistory of cardiac disease, to delay the onset, progression, or severityof the disease process(es).

In a preferred embodiment, the pharmaceutical composition isadministered to a person suffering from or at risk of developing organdamage, such as kidney disease or liver disease, to delay the onset,progression, or severity of the disease process(es).

In a preferred embodiment, the pharmaceutical composition isadministered to a person pre- and post-organ transplant to optimizetransplant success and avoid post-transplant complications associatedwith, for example, rejection and organ failure.

In a preferred embodiment, pharmaceutical composition is administered toan organ donor to preserve the organs before harvesting for transplant.

In a preferred embodiment, the pharmaceutical composition isadministered on a chronic basis to a normal or at-risk person for thepurpose of extending, prolonging, or maximizing lifespan potential.

In a preferred embodiment, the pharmaceutical composition isadministered on a chronic basis to canines, felines, and other non-humanmammal species for the purpose of extending, prolonging, or maximizinglifespan.

In a preferred embodiment, the olanzapine, risperidone, or reserpinecomponent of the combination exerts a body temperature-lowering effect.

Embodiments of the invention include pharmaceutical combinations ofolanzapine, risperidone, or reserpine with metformin.

Embodiments of the invention include pharmaceutical combinations ofolanzapine, risperidone, or reserpine with amlexanox.

Embodiments of the combination drug invention may include apharmaceutical component with temperature-reducing properties, such as aneurotensin analog, such as NT69L (Katz patent application US2012/0282227) or similar analogs, in combination with, for example,metformin or amlexanox.

Embodiments of the combination drug invention may include apharmaceutical component with body temperature-reducing properties, suchas β-blockers, clonidine, ethanol, phenothiazines, neuroleptics, andsedative hypnotics, with metformin or amlexanox.

EXAMPLES

Pharmaceutical Compositions

Pharmaceutical compositions of the present disclosure comprise at leastone compound, or tautomer, stereoisomer, pharmaceutically acceptablesalt or hydrate thereof, formulated together with one or morepharmaceutically acceptable carriers. These formulations include thosesuitable for oral, rectal, topical (e.g., drug-impregnated patch),buccal, and parenteral (e.g., subcutaneous, intramuscular, intradermal,or intravenous) administration. The most suitable form of administrationin any given case will depend on the degree and severity of thecondition being treated and on the nature of the particular compoundbeing used.

Formulations suitable for oral administration may be presented indiscrete units, such as capsules, cachets, lozenges, tablets, orpatches, each containing a predetermined amount of a compound of thepresent disclosure as powder or granules; as a solution or a suspensionin an aqueous or non-aqueous liquid; or as an oil-in-water orwater-in-oil emulsion. As indicated, such formulations may be preparedby any suitable method of pharmacy which includes the step of bringinginto association at least one compound of the present disclosure as theactive compound and a carrier or excipient (which may constitute one ormore accessory ingredients). The carrier must be acceptable in the senseof being compatible with the other ingredients of the formulation andmust not be deleterious to the recipient. The carrier may be a solid ora liquid, or both, and may be formulated with at least one compounddescribed herein as the active compound in a unit-dose formulation, forexample, a tablet, which may contain from ˜0.05% to ˜95% by weight ofthe at least one active compound. Other pharmacologically activesubstances may also be present including other compounds. Theformulations of the present disclosure may be prepared by any of thewell-known techniques of pharmacy consisting essentially of admixing thecomponents.

For solid compositions, conventional non-toxic solid carriers include,for example, pharmaceutical grades of mannitol, lactose, starch,magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose,magnesium carbonate, and the like. Liquid pharmacologicallyadministrable compositions can, for example, be prepared by dissolvingor dispersing, at least one active compound of the present disclosure asdescribed here and optional pharmaceutical adjuvants in an excipient,such as, for example, water, saline, aqueous dextrose, glycerol,ethanol, and the like, to thereby form a solution, ointment, orsuspension. Generally, suitable formulations may be prepared byuniformly and intimately admixing at least one active compound of thepresent disclosure with a liquid or finely divided solid carrier, orboth, and then, if necessary, shaping the product. For example, a tabletmay be prepared by compressing or molding a powder or granules of atleast one compound of the present disclosure, which may be optionallycombined with one or more accessory ingredients. Compressed tablets maybe prepared by compressing, in a suitable machine, at least one compoundof the present disclosure in a free-flowing form, such as a powder orgranules, which may be optionally mixed with a binder, lubricant, inertdiluent and/or surface active/dispersing agent(s). Molded tablets may bemade by molding, in a suitable machine, where the powdered form of atleast one compound of the present disclosure is moistened with an inertliquid diluent.

Formulations suitable for buccal (sub-lingual) administration includelozenges comprising at least one compound of the present disclosure in aflavored base, usually sucrose and acacia or tragacanth, and pastillescomprising the at least one compound in an inert base such as gelatinand glycerin or sucrose and acacia.

Formulations of the present disclosure suitable for parenteraladministration comprise sterile aqueous preparations of at least onecompound or tautomers, stereoisomers, pharmaceutically acceptable salts,and hydrates thereof, which are approximately isotonic with the blood ofthe intended recipient. These preparations are administeredintravenously, although administration may also be effected by means ofsubcutaneous, intramuscular, or intradermal injection. Such preparationsmay conveniently be prepared by admixing at least one compound describedherein with water and rendering the resulting solution sterile andisotonic with the blood. Injectable compositions according to thepresent disclosure may contain from ˜0.1 to ˜5% (w/w) of the activecompound.

Formulations suitable for rectal administration are presented asunit-dose suppositories. These may be prepared by admixing at least onecompound as described herein with one or more conventional solidcarriers, for example, cocoa butter, and then shaping the resultingmixture.

Formulations suitable for topical application to the skin may take theform of an ointment, cream, lotion, paste, gel, spray, aerosol, patch,or oil. Carriers and excipients that may be used include Vaseline,lanoline, polyethylene glycols, alcohols, and combinations of two ormore thereof. The active compounds are generally present at aconcentration of from ˜0.1% to ˜15% w/w of the composition, for example,from ˜0.5 to ˜2%.

The amount of active compound administered may be dependent on thesubject being treated, the subject's weight, the manner ofadministration and the judgment of the prescribing physician. Forexample, a dosing schedule may involve the daily or semi-dailyadministration of the encapsulated compound at a perceived dosage of ˜4μg to ˜1000 mg. In another embodiment, intermittent administration, suchas on a monthly or yearly basis, of a dose of the encapsulated compoundmay be employed. Encapsulation facilitates access to the site of actionand allows the administration of the active ingredients simultaneously,in theory producing a synergistic effect. In accordance with standarddosing regimens, physicians will readily determine optimum dosages andwill be able to readily modify administration to achieve such dosages.

A therapeutically effective amount of a compound or compositiondisclosed herein can be measured by the therapeutic effectiveness of thecompound. The dosages, however, may be varied depending upon therequirements of the patient, the severity of the condition beingtreated, and the compound being used. In one embodiment, thetherapeutically effective amount of a disclosed compound is sufficientto establish a maximal plasma concentration. Preliminary doses as, forexample, determined according to animal tests, and the scaling ofdosages for human administration is performed according to art-acceptedpractices.

Toxicity and therapeutic efficacy can be determined by standardpharmaceutical procedures in cell cultures or experimental animals,e.g., for determining the LD₅₀ (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD₅₀/ED₅₀.Compositions that exhibit large therapeutic indices are preferable.

Data obtained from the cell culture assays or animal studies can be usedin formulating a range of dosage for use in humans. Therapeuticallyeffective dosages achieved in one animal model may be converted for usein another animal, including humans, using conversion factors known inthe art (see, e.g., Freireich et al., Cancer Chemother. Reports50(4):219-244 (1966) and the following Table for Equivalent Surface AreaDosage Factors).

Approximate Equivalent Surface Area Dosage Factors

To: Mouse Rat Monkey Dog Human From: (20 g) (150 g) (3.5 kg) (8 kg) (60kg) Mouse 1 ½ ¼ ⅙   1/12 Rat 2 1 ½ ¼ 1/7 Monkey 4 2 1 ⅗ ⅓ Dog 6 4 ⅗ 1 ½Human 12 7 3 2 1

The dosage of such compounds lies preferably within a range ofcirculating concentrations that include the ED₅₀ with little or notoxicity. The dosage may vary within this range depending upon thedosage form employed and the route of administration utilized.Generally, a therapeutically effective amount may vary with thesubject's age, condition, and gender, as well as the severity of themedical condition in the subject. The dosage may be determined by aphysician and adjusted, as necessary, to suit observed effects of thetreatment.

Example 1

Mouse or Rat Experiments

Dose-ranging experiments of various combinations of the combination drugembodiment components are conducted to establish induction, range, andmaintenance of decreased body temperature in a healthy mouse or ratmodel.

In preliminary experiments, olanzapine, risperidone, and reserpine areevaluated independently at varying dose levels to determine the range ofbody temperature drop in mice or rats.

Such tests may also examine administration during the day versus eveningdosing to assess potential diurnal variation in body temperature. Thereis some evidence, for example, that olanzapine may be optimallyeffective in the ‘dark period’ with respect to temperature decrease.Metformin may be ideally suited for its desired effects occurring in the‘light period.’

Thus, combination testing is conducted to evaluate metformin with atleast one of olanzapine and risperidone and reserpine in ‘morning’ or‘evening’ combinations or with separate components being administered inthe morning and evening; for example, metformin in the morning andolanzapine in the evening, and assessing a single combinationchronotherapeutic formulation with delayed release of one component tomaximize diurnal characteristics/benefits of each drug's desired effect.

Example 2

Mouse or Rat Experiments

Separate 2-week dose-ranging and pharmacodynamic validation studies ofthree different drugs known to have temperature-lowering properties(olanzapine, risperidone, reserpine) are conducted in mice or rats toevaluate the optimal temperature-lowering pharmacodynamic effect anddose of each.

Of the three different drug candidates, two atypical anti-psychoticdrugs olanzapine and risperidone, and an indole alkaloid anti-psychoticand anti-hypertensive drug, reserpine, one is selected at theseexperiments for subsequent evaluation in combination with metformin, anoral anti-diabetic drug of the biguanide class.

As explained previously, metformin has caloric-restriction and otherdesired effects, such as weight loss, reduced insulin resistance, andincreased metabolism to balance or offset the potential for weight gain,decreased metabolism, and increased insulin resistance associated withanti-psychotics.

Each of the three drugs is first tested at doses at or near theequivalent therapeutic dose level for their approved indications (i.e.,translational equivalent mouse or rat dose as derived/calculated fromknown human dosing) followed by doses at one-half the translationaltherapeutic dose levels, followed by translational doses higher or lowerthan one-half the therapeutic dose, depending on outcome evaluations ateach level of testing. Parameters to be evaluated are as follows.

Primary objectives include:

-   -   Drop in temperature (actual and average/mean over time).    -   Sustainability of drop in temperature over time.    -   Intra-animal pattern of temperature drop.

Secondary objectives include:

-   -   Weight gain or loss.    -   Appetite.    -   Activity level.    -   Blood glucose levels.    -   Sleep efficacy

Example 3

Mouse or Rat Experiments

Based on an outcome evaluation of the Examples 1-2 experiments, one ofthe three drugs (and possibly an alternate) is designated as thecandidate(s) for the next stage, for evaluation as thetemperature-reducing component of a combination drug product.

In this example, the selected drug(s) and dose(s) fortemperature-lowering effect are evaluated when the drug is administeredin the morning or in the evening or as half-doses split between morningand evening.

Evaluation objectives are the same as those in Example 2, plus anevaluation of the temperature effect in the light and dark periods(i.e., a daytime and/or nighttime effect, regardless of when the drug(s)is(are) administered).

The combination with the most desirable overall profile is then selectedas the temperature-lowering component of the combination drug candidatefor further evaluation.

Example 4

Mouse or Rat Experiments

The selected temperature-lowering (anti-psychotic) drug candidate at theoptimal dose level is combined with three different doses of metforminat dose regimens and/or other combination permutations based on priorresults that may include one or more of the following:

-   -   Combination is dosed in the morning.    -   Combination is dosed in the evening.    -   Combination is split for morning and evening dosing.    -   Split drug-release regimens specific to diurnal variability (a        chronotherapeutic approach).    -   Other permutations as dictated by analyses of prior results.

Results of these are evaluated according to the objectives in Example 1,with particular emphasis on the offsetting impact of metformin on theundesired side effects of the temperature-lowering component of thecombination (the anti-psychotic).

Example 5

Non-Human Mammalian Progeria Model (e.g., Mouse or Rat)

The preferred combination drug is assessed, for example, in a mouseprogeria model. An established progeria mouse model experiment is usedto evaluate the effect of various combinations of the proposedcombination drug embodiment on lifespan in the progeria mouse (primaryendpoint) with secondary evaluations of weight and general health, aswell as necroscopy to evaluate the presence or stage of cardiovasculardisease, cerebral/neurological changes, stroke, tumors/cancer, organfunction.

Example 6

Rat Concussion Model

The preferred combination drug embodiment is assessed in an establishedexperimental model, such as the Wistar rat concussion model developed byViano et al. (2012) (also known as the National Football League Model;Ann. Blamed. Eng. 40, 213-226). The study measures (comparativeevaluations) parameters such as time to recovery, quality of recovery,extent of neurological damage, post-concussion sequelae presence andduration.

Example 7

Mouse or Rat Longevity Model

The preferred combination drug embodiment is assessed in a study similarin design to the Scripps study referenced earlier (Conti et al. (2006)Science 314, 825-828), to evaluate lifespan outcomes in treated andnon-treated animals.

TABLE 1 Experiments Primary Secondary Experimental Drug Doses EvaluationOutcome Outcome Overall Stage Evaluated Evaluated Period MeasuresMeasures Objective One* olanzapine Therapeutic dose, 2 weeks Drop intemp Weight gain Select one risperidone ½ therapeutic per drug (actualand or loss drug to reserpine dose, higher or per dose mean) appetiteadvance to lower than ½ Sustainability activity stage 2 therapeutic dose& patterns level for each drug. blood PD and dose- glucose rangingdesign. levels Mouse or rat. Two* Selected Mouse or rat full 2 weeks Asabove, Same as Select candidate dose morning; per plus stage 1 optimalfull dose Regimen light/dark dosing Half-dose split period effectpattern between morning & evening Mouse or rat Three Selected Threedifferent 2 weeks Same as Same as Proof-of- candidate metformin dosesper Stages one stage 1 concept, plus in combination Regimen and Twoselection of metformin with selected preferred Mouse or rat candidate.embodiment Possible regimen: drug in Morning dose; support of eveningdosing; IP claims split am/pm; chronotherapeutic dosing (TBD); other asdictated by prior results. Four A Preferred Mouse or rat TBD Effect ofOrgan Indication- combination progeria model lifespan damage; specificdrug cardio proof-of- embodiment disease concept progression Four BPreferred Wistar rat TBD Time to Comparative Indication- combinationconcussion model recovery extent of specific drug (NFL) neurologicalproof-of- embodiment damage; concept quality of recovery; post-concussion sequelae presence and duration Four C Combination Mouse ofrat TBD Longevity General Indication drug longevity model comparisonhealth POC

Embodiments of the present invention include at least the following:

A method for the pharmacological induction of a prolonged modestlyreduced body temperature (in the range of ˜95° F. to ˜98° F.) in apatient in need thereof.

A pharmaceutical composition comprising olanzapine and metformin thatreduces the body temperature (to the range of ˜95° F. to ˜98° F.) of ahuman patient in need thereof.

A chronotherapeutic pharmaceutical formulation comprising olanzapine andmetformin that safely lowers body temperature by BLSA study-like amounts(lower 50^(th) percentile), suitable for chronic low-dose use, providingeffective pharmacological means to achieve modest and controlledhypothermia (a decrease from normal baseline to the range of ˜95° F. to˜98° F.).

A pharmaceutical composition comprising doses of an atypicalanti-psychotic drug with modest hypothermia-inducing properties, such asolanzapine or risperidone or an indole alkaloid anti-psychotic andanti-hypertensive drug, such as reserpine, and an oral anti-diabeticdrug in the biguanide class, such as metformin.

A pharmaceutical composition comprising doses of an atypicalanti-psychotic drug with modest hypothermia-inducing properties, such asolanzapine or risperidone or an indole alkaloid anti-psychotic andanti-hypertensive drug, such as reserpine, and an anti-inflammatory,anti-allergic immunomodulatory drug, such as amlexanox.

A method of treating progeria by administering a pharmaceuticalcomposition comprising olanzapine and metformin

A method of treating progeria by administering a pharmaceuticalcomposition comprising risperidone and metformin.

A method of treating concussion using a pharmaceutical compositioncomprising reserpine and metformin.

A pharmaceutical composition comprising olanzapine, risperidone, orreserpine with metformin.

A pharmaceutical composition comprising olanzapine, risperidone, orreserpine with amlexanox.

A pharmaceutical composition comprising a neurotensin analog, such asNT69L, with metformin.

A pharmaceutical composition comprising a neurotensin analog, such asNT69L, with amlexanox.

A pharmaceutical composition comprising NT69L and metformin.

A pharmaceutical composition comprising NT69L and amlexanox.

A pharmaceutical composition comprising one component selected from thegroup consisting of β-blockers, clonidine, ethanol, phenothiazines,neuroleptics, and sedative hypnotics, with metformin.

A pharmaceutical composition comprising one component selected from thegroup consisting of I3-blockers, clonidine, ethanol, phenothiazines,neuroleptics, and sedative hypnotics, with amlexanox.

A method for the pharmacological induction of a prolonged, modestlyreduced body temperature (in the range of ˜95° F. to ˜98° F.) in apatient in need thereof.

A method for the pharmacological induction of a prolonged, modestlyreduced body temperature (in the range of ˜95° F. to ˜98° F.) in apatient in need thereof comprising administering a pharmaceuticalcomposition comprising metformin and a second agent, selected from thegroup consisting of olanzapine, risperidone, and reserpine.

A method for the pharmacological induction of a prolonged, modestlyreduced body temperature (in the range of ˜95° F. to ˜98° F.) in apatient in need thereof comprising administering a pharmaceuticalcomposition comprising metformin and olanzapine.

A method for the pharmacological induction of a prolonged, modestlyreduced body temperature (in the range of ˜95° F. to ˜98° F.) in apatient in need thereof comprising administering a pharmaceuticalcomposition comprising metformin and risperidone.

A method for the pharmacological induction of a prolonged, modestlyreduced body temperature (in the range of ˜95° F. to ˜98° F.) in apatient in need thereof comprising administering a pharmaceuticalcomposition comprising metformin and reserpine.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof comprisingdoses of an atypical anti-psychotic drug or an indole alkaloidanti-psychotic and anti-hypertensive drug, and an oral anti-diabeticbiguanide drug.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof comprisingdoses of an atypical anti-psychotic drug or an indole alkaloidanti-psychotic and anti-hypertensive drug, and an anti-inflammatory,anti-allergic immunomodulatory drug.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof comprisingmetformin and a second agent, selected from the group consisting ofolanzapine, risperidone, reserpine, and a neurotensin analog.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof comprisingamlexanox and a second agent, selected from the group consisting ofolanzapine, risperidone, reserpine, and a neurotensin analog.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof comprisingmetformin and NT69L.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof comprisingamlexanox and NT69L.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof comprisingmetformin and a second agent, selected from the group consisting ofβ-blockers, clonidine, ethanol, phenothiazines, neuroleptics, andsedative hypnotics.

A pharmaceutical composition that reduces the body temperature (to therange of ˜95° F. to ˜98° F.) of a patient in need thereof comprisingamlexanox and a second agent, selected from the group consisting ofβ-blockers, clonidine, ethanol, phenothiazines, neuroleptics, andsedative hypnotics.

A chronotherapeutic pharmaceutical formulation that reduces the bodytemperature (to the range of ˜95° F. to ˜98° F.) of a patient in needthereof.

A chronotherapeutic pharmaceutical formulation that reduces the bodytemperature (to the range of ˜95° F. to ˜98° F.) of a patient in needthereof comprising doses of an atypical anti-psychotic drug or an indolealkaloid anti-psychotic and anti-hypertensive drug, and an oralanti-diabetic biguanide drug.

A chronotherapeutic pharmaceutical formulation that reduces the bodytemperature (to the range of ˜95° F. to ˜98° F.) of a patient in needthereof comprising doses of an atypical anti-psychotic drug or an indolealkaloid anti-psychotic and anti-hypertensive drug, and ananti-inflammatory, anti-allergic immunomodulatory drug.

A chronotherapeutic pharmaceutical formulation that reduces the bodytemperature (to the range of ˜95° F. to ˜98° F.) of a patient in needthereof comprising metformin and a second agent, selected from the groupconsisting of P-blockers, clonidine, ethanol, phenothiazines,neuroleptics, and sedative hypnotics, with metformin.

A chronotherapeutic pharmaceutical formulation that reduces the bodytemperature (to the range of ˜95° F. to ˜98° F.) of a patient in needthereof comprising metformin and a second agent, selected from the groupconsisting of olanzapine, risperidone, reserpine, and a neurotensinanalog.

A chronotherapeutic pharmaceutical formulation that reduces the bodytemperature (to the range of ˜95° F. to ˜98° F.) of a patient in needthereof comprising amlexanox and a second agent, selected from the groupconsisting of olanzapine, risperidone, reserpine, and a neurotensinanalog.

All documents, publication, manuals, article, patents, summaries,references, and other materials cited here are incorporated by referencein their entirety. Other embodiments of the invention will be apparentto those skilled in the art from consideration of the specification andpractice of the invention disclosed here. It is intended that thespecification and examples be considered as exemplary, with the truescope and spirit of the invention indicated by the claims.

What is claimed is: 1-20. (canceled)
 21. A method of treating concussionin a patient in need thereof, said method comprising administering apharmaceutical composition comprising metformin and olanzapine.
 22. Themethod of claim 21, wherein said concussion is mild to moderate.
 23. Themethod of claim 22, wherein said concussion is mild.
 24. The method ofclaim 21, wherein said metformin and said olanzapine are administeredconcurrently.
 25. The method of claim 22, wherein said metformin andsaid olanzapine are administered concurrently.
 26. The method of claim23, wherein said metformin and said olanzapine are administeredconcurrently.
 27. The method of claim 21, comprising administeringeither: a) metformin in the morning and olanzapine in the evening; or b)olanzapine in the morning and metformin in the evening.
 28. The methodof claim 22, comprising administering either: a) metformin in themorning and olanzapine in the evening; or b) olanzapine in the morningand metformin in the evening.
 29. The method of claim 23, comprisingadministering either: a) metformin in the morning and olanzapine in theevening; or b) olanzapine in the morning and metformin in the evening.30. A method of ameliorating the neurological damage and/or symptomsassociated with concussion in a patient in need thereof, said methodcomprising administering a pharmaceutical composition comprisingmetformin and olanzapine.
 31. The method of claim 30, wherein saidconcussion is mild to moderate.
 32. The method of claim 31, wherein saidconcussion is mild.
 33. The method of claim 30, wherein said metforminand said olanzapine are administered concurrently.
 34. The method ofclaim 31, wherein said metformin and said olanzapine are administeredconcurrently.
 35. The method of claim 32, wherein said metformin andsaid olanzapine are administered concurrently.
 36. The method of claim30, comprising administering either: a) metformin in the morning andolanzapine in the evening; or b) olanzapine in the morning and metforminin the evening.
 37. The method of claim 31, comprising administeringeither: a) metformin in the morning and olanzapine in the evening; or b)olanzapine in the morning and metformin in the evening.
 38. The methodof claim 32, comprising administering either: a) metformin in themorning and olanzapine in the evening; or b) olanzapine in the morningand metformin in the evening.